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Background: Leishmania tarentolae, a unicellular eukaryotic protozoan, has been established as a novel host for recombinant protein production in recent years. Current protocols for protein expression in Leishmania are, however, time consuming and require extensive lab work in order to identify well-expressing cell lines. Here we established an alternative protein expression work-flow that employs recently engineered infrared fluorescence protein (IFP) as a suitable and easy-to-handle reporter protein for recombinant protein expression in Leishmania. As model proteins we tested three proteins from the plant Arabidopsis thaliana, including a NAC and a type-B ARR transcription factor. Results: IFP and IFP fusion proteins were expressed in Leishmania and rapidly detected in cells by deconvolution microscopy and in culture by infrared imaging of 96-well microtiter plates using small cell culture volumes (2 μL - 100 μL). Motility, shape and growth of Leishmania cells were not impaired by intracellular accumulation of IFP. In-cell detection of IFP and IFP fusion proteins was straightforward already at the beginning of the expression pipeline and thus allowed early pre-selection of well-expressing Leishmania clones. Furthermore, IFP fusion proteins retained infrared fluorescence after electrophoresis in denaturing SDS-polyacrylamide gels, allowing direct in-gel detection without the need to disassemble cast protein gels. Thus, parameters for scaling up protein production and streamlining purification routes can be easily optimized when employing IFP as reporter. Conclusions: Using IFP as biosensor we devised a protocol for rapid and convenient protein expression in Leishmania tarentolae. Our expression pipeline is superior to previously established methods in that it significantly reduces the hands-on-time and work load required for identifying well-expressing clones, refining protein production parameters and establishing purification protocols. The facile in-cell and in-gel detection tools built on IFP make Leishmania amenable for high-throughput expression of proteins from plant and animal sources.
We propose a paraconsistent declarative semantics of possibly inconsistent generalized logic programs which allows for arbitrary formulas in the body and in the head of a rule (i.e. does not depend on the presence of any specific connective, such as negation(-as-failure), nor on any specific syntax of rules). For consistent generalized logic programs this semantics coincides with the stable generated models introduced in [HW97], and for normal logic programs it yields the stable models in the sense of [GL88].
We reconsider the fundamental work of Fichtner ([2]) and exhibit the permanental structure of the ideal Bose gas again, using another approach which combines a characterization of infinitely divisible random measures (due to Kerstan,Kummer and Matthes [5, 6] and Mecke [8, 9]) with a decomposition of the moment measures into its factorial measures due to Krickeberg [4]. To be more precise, we exhibit the moment measures of all orders of the general ideal Bose gas in terms of certain path integrals. This representation can be considered as a point process analogue of the old idea of Symanzik [11] that local times and self-crossings of the Brownian motion can be used as a tool in quantum field theory. Behind the notion of a general ideal Bose gas there is a class of infinitely divisible point processes of all orders with a Levy-measure belonging to some large class of measures containing the one of the classical ideal Bose gas considered by Fichtner. It is well known that the calculation of moments of higher order of point processes are notoriously complicated. See for instance Krickeberg's calculations for the Poisson or the Cox process in [4].
A deterministic cycle scheduling of partitions at the operating system level is supposed for a multiprocessor system. In this paper, we propose a tool for generating such schedules. We use constraint based programming and develop methods and concepts for a combined interactive and automatic partition scheduling system. This paper is also devoted to basic methods and techniques for modeling and solving this partition scheduling problem. Initial application of our partition scheduling tool has proved successful and demonstrated the suitability of the methods used.
Afrika hat seinen berechtigten Platz auf der Weltbühne noch nicht gefunden. Ein wichtiger Grund dafür besteht darin, dass afrikanische Regierungen – im Gegensatz zu denen Chinas, Indiens, Brasiliens oder Indonesiens – nicht jene Maßnahmen ergriffen haben, die für das Entstehen eines Mittelstandes aus dem Gemenge von Millionen Armen wichtig sind.
I. Einleitung II. Ehepaare und eingetragene Lebenspartnerschaften III. Einstandsgemeinschaften IV. Die nicht funktionierende Bedarfsgemeinschaft V. Die vom Kind gebildete Bedarfsgemeinschaft VI. Stiefkinder in der Bedarfsgemeinschaft VII. Gemischte Bedarfsgemeinschaften VIII. Die zeitweise Bedarfsgemeinschaft IX. Haushaltsgemeinschaften
El artículo discute la formación de los conceptos de la naturaleza, espacio y morfología en la obra de Alexander von Humboldt y sus impactos en la constitución de la geografía física moderna. Influido por las reflexiones de Kant en la Crítica del Juicio, por los trabajos de Goethe y de Schelling, Humboldt desarrollará una nueva interpretación y representación de la naturaleza en la superficie terrestre, donde el concepto de espacialidad será fundamental para la explicación de los fenómenos de la naturaleza. La geografía física moderna se estructura a partir de un complejo entrecruzamiento de influencias, tanto estéticas como instrumentales trabajadas por Humboldt, donde el principio de la conexión será importante para la invención artística y científica del concepto de paisaje geográfico.
Alltagsorientierte Therapie
(2010)
I. Keine europäischen Vorgaben zur Haftung II. Anwendung allgemeiner Amtshaftungsgrundsätze III. System der Amtshaftung in Deutschland IV. Maßgebliche Amtshandlungen des Einheitlichen Ansprechpartners V. Maßgebliche Amtshandlungen zuständiger Stellen VI. Amtshaftung wegen fehlender elektronischer Erreichbarkeit? VII. Haftende Körperschaft